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Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology
Chinese hamster ovary (CHO) cells are the dominant mammalian host system for the production of recombinant therapeutic proteins. Improving the viable cell density during culture of recombinant CHO cells can greatly affect the production yield. MicroRNAs (miRs) -15a and 16-1 are known as negative reg...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Wolters Kluwer - Medknow
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6714117/ https://www.ncbi.nlm.nih.gov/pubmed/31516510 http://dx.doi.org/10.4103/1735-5362.263626 |
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author | Pairawan, Morvarid Sadat Bolhassani, Azam Rahimpour, Azam |
author_facet | Pairawan, Morvarid Sadat Bolhassani, Azam Rahimpour, Azam |
author_sort | Pairawan, Morvarid Sadat |
collection | PubMed |
description | Chinese hamster ovary (CHO) cells are the dominant mammalian host system for the production of recombinant therapeutic proteins. Improving the viable cell density during culture of recombinant CHO cells can greatly affect the production yield. MicroRNAs (miRs) -15a and 16-1 are known as negative regulators of multiple genes involved in cell cycle progression and apoptotic inhibition. miR sponges, which act as decoy targets, are transcripts which contain complementary binding sites to the seed region of related miRs. Stably expressed miR sponges are known as efficient tools for miR loss of function studies. In this study, stable CHO cell pools and clones expressing miRs-15a and 16-1 specific decoy transcript downstream of an enhanced green fluorescent protein reporter gene was developed. Analysis of cell growth during 12 days of batch culture indicated improved maximum viable cell density of CHO cells and clones expressing the decoy transcript. In addition, transient expression of a recombinant anti-CD52 monoclonal antibody was significantly improved in a decoy harboring CHO cell clone, representing a 3.37-fold increase in yield after 4 days of culture. Our results indicated that miR sponge technology can be successfully applied for the improvement of cell viability and transient monoclonal antibody expression in CHO host cells. |
format | Online Article Text |
id | pubmed-6714117 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Wolters Kluwer - Medknow |
record_format | MEDLINE/PubMed |
spelling | pubmed-67141172019-09-12 Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology Pairawan, Morvarid Sadat Bolhassani, Azam Rahimpour, Azam Res Pharm Sci Original Article Chinese hamster ovary (CHO) cells are the dominant mammalian host system for the production of recombinant therapeutic proteins. Improving the viable cell density during culture of recombinant CHO cells can greatly affect the production yield. MicroRNAs (miRs) -15a and 16-1 are known as negative regulators of multiple genes involved in cell cycle progression and apoptotic inhibition. miR sponges, which act as decoy targets, are transcripts which contain complementary binding sites to the seed region of related miRs. Stably expressed miR sponges are known as efficient tools for miR loss of function studies. In this study, stable CHO cell pools and clones expressing miRs-15a and 16-1 specific decoy transcript downstream of an enhanced green fluorescent protein reporter gene was developed. Analysis of cell growth during 12 days of batch culture indicated improved maximum viable cell density of CHO cells and clones expressing the decoy transcript. In addition, transient expression of a recombinant anti-CD52 monoclonal antibody was significantly improved in a decoy harboring CHO cell clone, representing a 3.37-fold increase in yield after 4 days of culture. Our results indicated that miR sponge technology can be successfully applied for the improvement of cell viability and transient monoclonal antibody expression in CHO host cells. Wolters Kluwer - Medknow 2019-08 /pmc/articles/PMC6714117/ /pubmed/31516510 http://dx.doi.org/10.4103/1735-5362.263626 Text en Copyright: © 2019 Research in Pharmaceutical Sciences http://creativecommons.org/licenses/by-nc-sa/4.0 This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms. |
spellingShingle | Original Article Pairawan, Morvarid Sadat Bolhassani, Azam Rahimpour, Azam Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title | Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title_full | Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title_fullStr | Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title_full_unstemmed | Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title_short | Enhanced transient expression of an anti-CD52 monoclonal antibody in CHO cells through utilization of miRNA sponge technology |
title_sort | enhanced transient expression of an anti-cd52 monoclonal antibody in cho cells through utilization of mirna sponge technology |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6714117/ https://www.ncbi.nlm.nih.gov/pubmed/31516510 http://dx.doi.org/10.4103/1735-5362.263626 |
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